CN105838636B - A method of improving lactic acid bacteria biomass under high salt conditions - Google Patents
A method of improving lactic acid bacteria biomass under high salt conditions Download PDFInfo
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- CN105838636B CN105838636B CN201510871124.1A CN201510871124A CN105838636B CN 105838636 B CN105838636 B CN 105838636B CN 201510871124 A CN201510871124 A CN 201510871124A CN 105838636 B CN105838636 B CN 105838636B
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Abstract
The invention discloses a kind of methods for improving lactic acid bacteria biomass under high salt conditions, belong to technical field of bioengineering.The present invention is will to pass through the seed liquor of 4 DEG C of Cold pretreatments, accesses in the fermentation medium containing 12%NaCl and cultivates 96 hours, biomass increases than the cell without Cold pretreatment.The method of the present invention is simple, is convenient for operation, can be used for the fermenting and producing of lactic acid bacteria.
Description
Technical field
The present invention relates to a kind of methods for improving lactic acid bacteria biomass under high salt conditions, belong to biotechnology neck
Domain.
Background technique
Thermophilic salt tetrads (Tetragenococcus halophilus) belongs to the Tetracoccus in lactic acid Cordycepps, extensively
It is general to be present in the salting food products such as soy sauce, bean cotyledon and alec.Thermophilic salt tetrads makes the color of these traditional fermented foods more
Light covers food saline taste, promotes the generation of flavor component, play an important role to the flavor of fermented seasonings.But it is passing
It unites in fermented product, hypersaline environment seriously inhibits the growth performance and metabolic activity of thallus, the physiological activity of cell is affected,
It is severely impacted the quality of product.Therefore, survival rate of the thermophilic salt tetrads under high salt conditions is improved with important meaning
Justice.
The application of thermophilic salt tetrads in the food industry, the stressful environmental suffered from are multiple and complicated.It adapts to
Property reaction be the common one kind of lactic acid bacteria stress protected mode, when cell is in stressful environmental, by a kind of adaptation reaction
The protective effect induced can protect cell and resist more rugged environment stress, these reactions can be used for improving lactic acid
Survival of the bacterium under stress conditions.
Summary of the invention
The object of the present invention is to provide a kind of sides that lactic acid bacteria biomass under high salt conditions is improved using Cold pretreatment
Method.
The Cold pretreatment condition is 4 DEG C, 5-15 h.
Bacterial strain: thermophilic salt tetrads (Tetragenococcus halophilus) CGMCC 3792, laboratory is from soy sauce
It is separated in wine with dregs, is sequenced and identifies through form, biophysical and biochemical tests and 16S rDNA, be preserved in Chinese microorganism strain preservation pipe
Reason committee common micro-organisms center (China General Microbiological Culture
CollectionCenter, CGMCC), depositary institution address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences
Institute of microbiology；The deposit date is on April 29th, 2010, deposit number was CGMCC NO.3792, and classification naming is thermophilic salt
Tetrads (Tetragenococcus halophilus).
Seed culture method: seed culture medium be 5 g/L tryptones, 5 g/L soy peptones, 5 g/L beef extracts,
2.5 g/L yeast extracts, 0.5 g/L ascorbic acid, 0.25 g/L MgSO4.7H2O, 19 g/L sodium β-glycerophosphates, pH 7.0.
The lactic acid bacteria glycerol tube storing liquid of -80 DEG C of preservations is taken to be inoculated in seed culture medium with the inoculum concentration of 5% (V/V), 30 DEG C of standings
Cultivate 24 h.
Cold pretreatment method: by cultured seed liquor with 10000 r/min(4 DEG C) 5 min of centrifugation collect thallus, and it will
Thallus is resuspended in 4 DEG C of Cold pretreatment 5-15 h of progress in fresh seed culture medium.
High salt fermentation cultural method: high salt fermentation culture medium is 5 g/L tryptones, 5 g/L soy peptones, 5 g/L
Beef extract, 2.5 g/L yeast extracts, 0.5 g/L ascorbic acid, 0.25 g/L MgSO4.7H2O, 19 g/L sodium β-glycerophosphates,
NaCl concentration is adjusted to as 12%, pH 7.0.Take by the pretreated seed liquor of low temperature with 5%(V/V) inoculum concentration be inoculated in it is with high salt
In fermentation medium, 30 DEG C of 96 h of stationary culture.
The measurement of biomass: thalline were collected by centrifugation for the bacteria suspension for taking after fermentation, suitable with diluting after sterile water washing 3 times
When multiple, shake up, with TU-1901 two-beam light ultraviolet-uisible spectrophotometer, the size of turbidimetric assay OD value at 600 nm.
Beneficial effects of the present invention: the kind of 4 DEG C of Cold pretreatments is passed through in access in the fermentation medium provided by the invention
Sub- liquid, thermophilic salt tetrads CGMCC NO.3792 are (lower without 4 DEG C than control group under (12%NaCl) stress conditions with high salt
Temperature pretreatment) there is better growth performance.The present invention provides a kind of based on the thermophilic salt tetrads salt side of body of Cold pretreatment promotion
Compel the method for resistance, it is simple and easy, the collaboration for starting adaptation mechanism when lactic acid bacteria copes with extraneous Different stress environment can be explored
The mode of action.
Specific embodiment
Example 1: seed liquor is without 4 DEG C of Cold pretreatments (control group), the life of T.halophilusCGMCC NO.3792
Long situation (table 1)
The glycerin storage liquid of -80 DEG C of preservations is taken to be inoculated in seed culture medium with 5% inoculum concentration, 30 DEG C of stationary cultures are for 24 hours
To logarithm middle and later periods (hereinafter referred to as seed liquor).By r/min(4 DEG C of seed liquor 10000) 5 min of centrifugation collect thallus, by thallus
It is resuspended in fresh seed culture medium, is seeded in high salt fermentation culture medium with 5% inoculum concentration and cultivates 96 h, measure OD600
Value is 2.03.
Example 2: seed liquor passes through 4 DEG C of Cold pretreatment 5-15 h(experimental groups), T.halophilusCGMCC NO.3792
Growing state (table 1)
By r/min(4 DEG C of seed liquor 10000) 5 min of centrifugation collect thallus, thallus is resuspended in fresh seed culture
After carrying out 4 DEG C of Cold pretreatments 5 h, 10 h and 15 h in base respectively, it is seeded in high salt fermentation culture medium with 5% inoculum concentration
96 h are cultivated, OD is measured600Value is respectively 2.26,2.20 and 2.86, and biomass has been respectively increased 11.3% compared with the control group,
8.4% and 40.9%.
Fungal biodiversity in 1 high salt fermentation culture medium of table
Claims (5)
1. a kind of method for improving lactic acid bacteria biomass under high salt conditions comprising the steps of:
(1) a kind of lactobacillus solution of Cold pretreatment；Wherein, the treatment temperature of Cold pretreatment is 4 DEG C；
(2) lactobacillus solution is accessed in a kind of high salt fermentation culture medium and is cultivated；
The lactic acid bacteria is the thermophilic salt tetrads (Tetragenococcus that deposit number is CGMCC NO.3792
Halophilus).
2. improving the method for lactic acid bacteria biomass under high salt conditions according to claim 1, it is characterised in that: the low temperature
Pretreatment time is 5 hours to 15 hours.
3. improving the method for lactic acid bacteria biomass under high salt conditions according to claim 1, it is characterised in that: the lactic acid
The inoculum concentration of bacterium is calculated as 5% by percent by volume.
4. improving the method for lactic acid bacteria biomass under high salt conditions according to claim 1, it is characterised in that: the lactic acid
The preparation of bacterium seed liquor and Cold pretreatment comprising the steps of:
(1) seed culture medium are as follows: 5 g/L tryptones, 5 g/L soy peptones, 5 g/L beef extracts, 2.5 g/L yeast extracts,
0.5 g/L ascorbic acid, 0.25 g/LMgSO4.7H2O, 19 g/L sodium β-glycerophosphates, pH 7.0；
(2) the lactic acid bacteria glycerol tube storing liquid of -80 DEG C of preservations is taken, the lactic acid bacteria is inoculated with the inoculum concentration that percent by volume is 5%
Glycerol tube storing liquid is in seed culture medium, 30 DEG C of 24 h of stationary culture；
(3) seed liquor is collected into thallus with 10000 r/min, 4 DEG C of 5 min of centrifugation, thallus is resuspended in fresh seed culture
In base, pre-processed under the conditions of 4 DEG C.
5. improving the method for lactic acid bacteria biomass under high salt conditions according to claim 1, it is characterised in that: described with high salt
Fermentation medium preparation and condition of culture comprising the steps of:
(1) high salt fermentation culture medium are as follows: 5 g/L tryptones, 5 g/L soy peptones, 5 g/L beef extracts, 2.5 g/L yeast
Cream, 0.5 g/L ascorbic acid, 0.25 g/LMgSO4·7H2O, 19 g/L sodium β-glycerophosphates, NaCl concentration are adjusted to 12%, pH
It is 7.0；
(2) condition of culture are as follows: the inoculum concentration for being 5% with percent by volume is inoculated with the seed liquor in high salt fermentation culture medium, and 30
DEG C 96 h of stationary culture.
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| CN107087645A (en) * | 2017-04-01 | 2017-08-25 | 杨飞 | A kind of biological mixed pesticide of pest control, preparation method and application |
| CN109182244B (en) * | 2018-08-01 | 2021-11-02 | 天津大学 | Method for improving biomass of bacillus natto under high-temperature condition |
| CN109370933B (en) * | 2018-09-28 | 2021-11-02 | 四川大学 | Co-culture method for improving acid resistance of saccharomycetes |
| CN111662860A (en) * | 2020-07-22 | 2020-09-15 | 四川大学 | Method for improving survival rate of tetragenococcus halophilus under extreme conditions |
| CN113637626B (en) * | 2021-09-27 | 2023-07-11 | 宜宾五粮液股份有限公司 | Method for improving survival rate of Russell yeast under high-salt stress condition |
| CN114561328A (en) * | 2022-04-18 | 2022-05-31 | 四川大学 | Preparation method of high-activity lactobacillus agent |
| CN115181769B (en) * | 2022-08-24 | 2023-08-08 | 四川大学 | Method for improving yield of extracellular polysaccharide of tetracoccus halophilus |
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| Characterization of a Multiple-stress Tolerance Tetragenococcus halophilus and Application as Starter Culture in Chinese Horsebean-Chili-Paste Manufacture for Quality Improvement;Chongde Wu等;《Food Sci. Technol. Res.》;20131231;第19卷(第5期);第855-864页 * |
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